US10905128B2 - Trichoderma compositions and methods of using the same - Google Patents
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- US10905128B2 US10905128B2 US16/270,834 US201916270834A US10905128B2 US 10905128 B2 US10905128 B2 US 10905128B2 US 201916270834 A US201916270834 A US 201916270834A US 10905128 B2 US10905128 B2 US 10905128B2
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungi isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/38—Trichoderma
-
- C12R1/885—
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
Definitions
- Blast and sheath blight diseases of rice as well as other plant diseases are caused by two known fungal pathogens, namely Magnaporthe oryzae and Rhizoctonia solani . They are widespread and economically important problems of rice production in southern rice producing states of the USA and worldwide. Historically, these two diseases have been the most common biological causes of yield loss in Arkansas and some popular higher yielding rice varieties have been removed from production due in particular to blast disease. Moreover, development of resistance to current commercial fungicides is of great concern. A large acreage of rice is sprayed with fungicides every year. Besides, use of chemicals for crop diseases suppression is environmentally unsafe.
- BCAs Biocontrol Agents
- Trichoderma Species of Trichoderma have been shown to provide varying levels of biological control in a number of important plant pathogens, including Sclerotium cepivorum (Abd-el Moity et al, 1983, Abd-el Moity et al, 1981), Armillaria mellea (Ohr et al, 1973), Rhizoctonia solani (Chet et al, 1981, Elad et al, 1982, Ruppel et al, 1983), Verticillium dahliae (Jordan et al, 1978, Marios et al, 1982) and Pythium spp.
- Sclerotium cepivorum Abd-el Moity et al, 1983, Abd-el Moity et al, 1981
- Armillaria mellea Ohr et al, 1973
- Rhizoctonia solani Rhizoctonia solani
- Verticillium dahliae Verticillium dahl
- Trichoderma -plant interactions are complex. An important fact is that different strains substantially vary in their effects on plants. Trichoderma strains have been already used as biocontrol agents against numerous plant pathogens and quite a few have been developed for use in commercial production.
- Trichoderma harzianum known as Trichodex®
- Trichodex® Trichoderma harzianum
- Ecochodex® Trichodex®
- Ecochodex® Trichoderma harzianum
- BCAs BCAs that can be used to control devastating plant diseases caused by, for example, Magnaporthe oryzae and Rhizoctonia solani.
- a new Trichoderma strain designated as TM17 and deposited under accession number NRRL 67984 is provided.
- the present invention relates to agricultural compositions.
- the agricultural compositions may include Trichoderma strain TM17 and a carrier.
- plants are provided.
- the plants may include Trichoderma strain TM17 or any one of the agricultural compositions described herein.
- the Trichoderma strain TM17 or any one of the agricultural compositions described herein may be present on or within at least a part of the plant.
- methods for inhibiting the growth of a microorganism on a plant may include contacting the plant with an effective amount of Trichoderma strain TM17 or any one of the agricultural compositions described herein to inhibit the growth of the microorganism on the plant.
- the plant may be a rice plant and the microorganism may be a fungi.
- the plant may be contacted by foliage spray application of the agricultural compositions or the Trichoderma strain TM17.
- FIG. 1 shows the methodology used to isolate, characterize and the final identification protocol for the endophyte Trichoderma sp. TM17.
- FIGS. 2A-2D show the dual confrontation result of TM17 against Magnaporthe oryzae
- FIGS. 2A, 2B, 2C, and 2D are day 3, 4, 6, and 10, respectively.
- BL-Blast pathogen Magnaporthe oryzae
- TR Trichoderma sp.-TM17
- TR+BL dual plate for TR and BL in a single plate.
- FIGS. 3A-3C show the dual confrontation result of TM17 against Rhizoctonia solani AG1-1A.
- FIGS. 3A, 3B, 3C are day 3, 4, 6, respectively.
- SB-Sheath blight Rhizoctonia solani AG1-1A
- TR- Trichoderma sp.-(TM17) and TR+SB dual plate for TR and SB in a single plate.
- FIG. 4 shows the growth performance of BL, SB, TR and dual inoculated plates after 10 days of inoculation.
- FIGS. 5A-5D show the performance of rice plants treated in accordance with the present invention.
- FIG. 5A shows control treatment that was inoculated with the blast pathogen but with no TM17 applied.
- FIG. 5B shows plants pre-treated with TM17 and then inoculated with the blast pathogen.
- FIGS. 5C and 5D show inoculated plants but the healthy looking plants on the left are treated with TM17 prior to inoculation with the blast pathogen.
- FIG. 6 shows the visible lesion size on the rice leaves.
- the top picture shows the results when TM17 is sprayed 24 h before inoculation with the blast pathogen.
- the middle picture shows the results of post-treatment with TM17.
- the TM17 was applied 48 h after the blast pathogen inoculation.
- the picture at the bottom shows the control plants inoculated with the blast pathogen and not treated with TM17.
- FIGS. 7A and 7B show two photographs to demonstrate the contrast between TM17 treated and untreated rice plants from two different views.
- the plants on the left of each picture were inoculated with Rhizoctonia solani AG1-1A agar plugs 24 h after TM17 treatment and the plants on the right were inoculated with Rhizoctonia solani AG1-1A agar plugs, but not treated with TM17.
- TM17 Trichoderma strain
- spraying plants with TM17 allows TM17 to colonize the plant surface thereby creating a plant- Trichoderma interaction system.
- the plant- Trichoderma interaction system protects the plants from being colonized by, and damaged by, pathogenic microbes such as fungi.
- TM17 may provide a broad-spectrum control of multiple diseases in plants and provide a safe alternative to chemical pesticides.
- TM17 a new Trichoderma strain designated as TM17 is provided.
- a viable culture of TM17 has been deposited in the ARS Culture Collection (NRRL), Peoria, Ill. under the accession number NRRL 67984.
- Trichoderma strain TM17 produces abundant spores and has been shown to suppress and control the rice blast and sheath blight diseases caused by Magnaporthe oryzae and Rhizoctonia solani AG1-1A, respectively.
- the present invention relates to a method of protecting plants from infection, which comprises contacting the plant with Trichoderma strain TM17 under conditions effective for the Trichoderma strain to colonize the surface of the plant, thereby creating a plant- Trichoderma interaction system.
- a biologically pure culture of fungus Trichoderma sp. isolate TM17 for inducing suppression of rice plant diseases caused by Magnaporthe oryzae and Rhizoctonia solani AG1-1A is provided.
- the culture produces abundant mycelium, conidia and chlamydospores.
- an “agricultural composition” is a composition formulated for application to a plant or plant part.
- An agricultural composition is commonly in liquid form for application by spraying or soaking, but may be in a solid, granular, or powder form for rehydration or application by dusting or dry coating.
- the agricultural composition may be concentrated for dilution in water or other solvent.
- the agricultural compositions may include Trichoderma strain TM17 and a carrier.
- a “carrier” may be solid or liquid and may include substances ordinarily employed in formulations applied to plants.
- Carriers may include a buffer, water, oil, nonionic surfactants, ionic surfactants such as cationic or anionic surfactants, or available agricultural byproducts from, for example and without limitation, rice.
- the agricultural compositions may also include an additional active ingredient such as, without limitation, a fungicide, an herbicide, a biosanitizer product or fertilizer.
- an additional active ingredient such as, without limitation, a fungicide, an herbicide, a biosanitizer product or fertilizer.
- the agricultural compositions may include Trichoderma strain TM17 at a concentration between 10 5 to 10 12 or more conidia per milliliter or any range therein.
- the concentration of the Trichoderma strain TM17 in the agricultural composition may be 10 7 to 10 10 conidia per milliliter.
- the concentration of Trichoderma strain TM17 conidia in the agricultural composition may be quantified using a hemocytometer.
- plants are provided.
- the plants may include Trichoderma strain TM17 or any one of the agricultural compositions described herein.
- the Trichoderma strain TM17 or any one of the agricultural compositions described herein may be present on or within at least a part of the plant.
- a “plant” includes any portion of the plant including, without limitation, a whole plant or a portion of a plant such as a part of a root, leaf, stem, seed, pod, flower, cell, tissue plant germplasm, asexual propagate, or any progeny thereof.
- a rice plant refers to the whole rice plant or portions thereof including, without limitation, the leaves, roots, or otherwise.
- Suitable “plants” may include, without limitation, a rice plant, a cotton plant, a soybean plant, a wheat plant, a sorghum plant, or a corn plant.
- the plant is a rice plant.
- methods for inhibiting the growth of a microorganism on a plant are provided.
- the methods may include contacting the plant with an effective amount of Trichoderma strain TM17 or any one of the agricultural compositions described herein to inhibit the growth of the microorganism on the plant.
- the “microorganism” may be any bacterial or fungal plant pathogen.
- the microorganism may be a fungal plant pathogen including, without limitation, a Magnaporthe species or a Rhizoctonia species.
- the microorganism is Magnaporthe oryzae or Rhizoctonia solani .
- the Rhizoctonia solani may be Rhizoctonia solani AG1-1A.
- Magnaporthe oryzae is the causative agent of blast disease in rice and Rhizoctonia solani AG1-1A is the causative agent of sheath blight.
- contacting may be carried out through any of the variety of procedures used to apply compositions to plants that will be apparent to the skilled artisan. Suitable application methods may include, without limitation spraying or dusting. Other suitable application procedures can be envisioned by those skilled in the art. Contacting may also be carried out indirectly via application, for example, to the soil surrounding a plant, trunk injection, or other plant media or substrates. The “contacting” of the present methods may be carried out before or after the microorganism grows on the plant.
- various parts of the plant may be contacted with or by Trichoderma strain TM17 or any one of the agricultural compositions described herein.
- the leaves or seeds of the plant may be contacted with Trichoderma strain TM17 or any one of the agricultural compositions described herein.
- the plant may be contacted at least 2, 3, 4, 5, or more times with with Trichoderma strain TM17 or any one of the agricultural compositions described herein during a single growing season. Different parts of the plant may be contacted at different points within the growing season. The plant may be contacted with the Trichoderma strain TM17 or the agricultural compositions using different formulations or means of contacting at different points within a growing season.
- Effective amount is intended to mean an amount of a composition described herein sufficient to inhibit the growth of a microorganism on a plant by, for example, 10%, 20%, 50%, 75%, 80%, 90%, 95%, or 1-fold, 3-fold, 5-fold, 10-fold, 20-fold, or more compared to a negative control plant not treated with the Trichoderma strain TM17 or one of the agricultural compositions provided herein.
- the effective amount of Trichoderma strain TM17 either alone or in an agricultural composition may be 10 5 to 10 12 or more conidia per milliliter or any range therein.
- the concentration of Trichoderma strain TM17 either alone or in an agricultural composition is 10 7 to 10 10 conidia per milliliter.
- a “negative control” refers to a sample that serves as a reference for comparison to a test sample.
- a test sample can be taken from a test condition including the presence of Trichoderma strain TM17 and compared to negative control samples lacking Trichoderma strain TM17 or including a composition not expected to inhibit microbial growth.
- controls can be designed for assessment of any number of parameters.
- RNA Unless otherwise specified or indicated by context, the terms “a”, “an”, and “the” mean “one or more.”
- a protein or “an RNA” should be interpreted to mean “one or more proteins” or “one or more RNAs,” respectively.
- Trichoderma sp.-TM17 is the first microorganism isolated from rice seed as an endophyte that showed a potential to control both Blast and Sheath blight disease of rice in Arkansas.
- Trichoderma sp.-TM17 The Trichoderma strain disclosed herein, Trichoderma sp.-TM17, was isolated from rice seed as an endophyte. The isolation of the endophytic strain was accomplished by the following procedure. See, e.g., FIG. 1 . Rice seeds were collected from different sites of major rice growing fields of Arkansas at rice crop maturity and were kept in paper bags at 4° C. until use. Seed samples randomly selected from the mixed samples were set for processing for the test. Isolation of endophyte fungi was according to the protocol of Mysore et al. 2005. Briefly, seeds were cleaned under running tap water to remove debris and then air-dried on a sterile filter paper.
- Conidia and mycelia of TM17 were produced by growing the isolate on PDA agar for 7 days under continuous fluorescent light at room temperature. Conidia were removed from the agar surface by pipetting 5 ml of sterile distilled water on the surface and gently rubbing the surface with a sterile cotton-tipped applicator. Conidia were counted in a hemocytometer, and the suspensions adjusted with water to provide the desired concentration of conidia in each test. There were five replications in all experiments, and each experiment was done twice.
- Biocontrol activities of Trichoderma sp. strain TM17 against the two fungal rice pathogens were determined in vitro on potato dextrose agar (PDA) (20 ml/plate). See FIGS. 2A-2D, 3A-3C, and 4 .
- the two pathogens were Magnaporthe oryzae ( FIGS. 2A-2D and 4 ) and Rhizoctonia solani AG1-1A ( FIGS. 3A-3C and 4 ).
- a plug of the biocontrol agent (8 mm diameter) from the edge of 5-day old colonies of the Trichoderma cultures and of 10-day old colonies of the pathogens were paired on the medium on opposite sides of 9 ⁇ 1.5 cm petri plates and then were incubated at room temperature.
- Antagonistic potential presented as averaged growth inhibition or overgrowth of each plant pathogen (prey) by Trichoderma strain TM17 and was expressed in percentages. In both cases 100% represented unrestricted growth of the prey corrected for the confrontation with itself.
- Trichoderma sp.-TM17 showed an aggressive biocontrol potential growing faster and eventually overwhelming the tested pathogen isolates compared to the control plates inoculated with the respective pathogen alone under the same conditions of incubation.
- the Trichoderma sp.-TM17 did not produce zones of inhibition against any of the pathogens on plates. However, its competitive growth overwhelmed the pathogen growth. See FIGS. 2A-2D, 3A-3C, and 4 .
- Trichoderma sp.-TM17 The ability of Trichoderma sp.-TM17 to suppress diseases was also determined in vivo. This endophyte Trichoderma isolate revealed high inhibition potential on the growth of the two prey isolates. Greenhouse experiments were carried out for both blast ( FIGS. 5A-5D , FIG. 6 ) and sheath blight ( FIGS. 7A-7B ) control by TM17. To test for blast, seeds of M2O6 planted in hill plots in a 0.02 m 2 flat containing greenhouse soil:sand:loam mixture in 3:1:1 proportion, respectively. There were 81 hill plots in a flat. The flats were kept in the greenhouse at an average temperature of 78° F. and 85% relative humidity. Flats were watered for 3 weeks and fertilized as needed.
- Conidia of TM17 harvested from 8 to 10-day-old cultures growing on PDA were washed off using cotton-tipped applicator. Suspensions of conidia were counted using a hemocytometer and adjusted to contain 10 9 conidia per milliliter for spray application.
- TM17 was sprayed on rice seedlings at full tillering developmental stage and then after 24 hours the plants were inoculated following standard greenhouse procedure by placing 10 days old mycelial agar plugs of Rhizoctonia solani AG1-1A at the base of each culm.
- TM17 was sprayed 24 hours after inoculation at a concentration of 10 9 conidia per ml.
- a conidial suspension of M oryzae was prepared by growing it on rice bran agar at 27° C. for 10 days, washed with sterilized distilled water and strained through sterile filter papers to remove mycelia. Conidial concentrations of M oryzae were adjusted to 10 5 conidia per ml. Then the suspensions were kept at 4° C. until foliage spray.
- TM17 was sprayed on rice seedlings at 24 h before spray-inoculation by M oryzae suspension.
- rice seedlings of similar age were spray-inoculated with the spore suspension of M oryzae at 24 h before spraying with TM17 at 10 9 conidia per ml.
- Trichoderma sp.-TM17 Application of Trichoderma sp.-TM17:
- TM17 Conidial suspension of TM17 was prepared for protective and therapeutic treatments.
- TM17 was sprayed 48 hours before inoculating the respective pathogens ( FIGS. 5B-5C ).
- rice plants were inoculated with isolates of Magnaporthe oryzae and Rhizoctonia solani AG1-1A, then 48 h after the rice plants were sprayed with TM17 suspension ( FIGS. 5A-5B ).
- Experimental controls were treated with water in place of TM17.
- Trichoderma sp.-TM17 which is seen potentially useful in controlling the two major rice diseases caused by Magnaporthe oryzae and Rhizoctonia solani AG1-1A.
- TM17 a strain of Trichoderma sp. was applied to greenhouse grown rice plants in an amount sufficient to colonize and populate the plant surface, thereby reducing/suppressing the growth of the pathogens. See FIGS. 5A-5D , FIG. 6 , and FIG. 7A-7B .
- TM17 A deposit of the University of Arkansas Division of Agriculture proprietary Trichoderma sp. strain designated as TM17 (RREC F21) disclosed above and recited in the appended claims has been made with the ARS Culture Collection (NRRL), 1815 N. University Street, Peoria, Ill. 61604. The date of deposit was Sep. 18, 2020. The deposit comprises 5 liquid nitrogen stocks, which were found viable on Sep. 18, 2020. All restrictions will be irrevocably removed upon granting of a patent, and the deposit is intended to meet all of the requirements of 37 C.F.R. ⁇ 1.801-1.809. The NRRL Accession Number is 67984. The deposit will be maintained in the depository for a period of thirty years, or five years after the last request, or for the enforceable life of the patent, whichever is longer, and will be replaced as necessary during that period.
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Abstract
Description
- Abd-El-Moity T H, Shatla M N, 1981. Biological control of white rot disease of onion (Sclerotium cepivorum) by Trichoderma harzianum. Phytopathologische Zeitshrift 100, 29-35.
- Abd-El-Moity T H, Papavizas G C, Shatla M N, 1983. Induction of new isolates of Trichoderma harzianum tolerant to © 2002 BSPP Plant Pathology (2002) 51, 735-745 744 J. P. Clarkson et al. fungicides and their experimental use for control of white rot of onion. Phytopathology 72, 396-400.
- Benitez, T., A. M. Rincon, M. C. Limon and A. C. Codon. 2004. Biocontrol mechanism of Trichoderma strains. International Microbiol., 7: 249-260.
- CHET. I. & BAKER, P., 1981. Isolation and biocontrol potential of Trichoderma hamatum from soil naturally suppressive of Rhizoctonia solani. Phytopathology 71, 286-290.
- Cook R, Baker K F. 1983. The nature and practice of biological control of plant pathogens. St Paul, M N, USA: The American Phytopathological Society.
- Elad Y. 2000. Trichoderma harzianum T39 preparation for biocontrol of plant disease-control of Botrytis cinerea, Sclerotinia sclerotiorum and Cladosporium flavum. Biocontrol Sci Techn; 10: 499-507.
- Elad, Y., Kalfon, A. and Chet, I. 1982. Control of Rhizoctonia solani in cotton by seed-coating with Trichoderma spp. spores. Plant Soil 66: 279-281.
- Hadar Y, Harman G E, Taylor A G. 1984. Evaluation of Trichoderma Koningii and T hamatum from New York for biological control of seed rot caused by Pythium spp. Phytopathology, 74: 106-110.
- Harman G E. 2000. Myths and dogmas of biocontrol: changes in perceptions derived from research on Trichoderma harzianum T-22. Plant Dis; 84(4): 377-393.
- Harman, G. E. and Hadar, Y. 1983. Biological control of Pythium species. Seed Sci. Technol. 11:893-906.
- Marois J. J., S. A. Jahnson, M. T. Dunn and G. C. Papavizas. 1982. Biological control of Verticillium wilt of eggplant in the field. Plant Disease 66, 1166-1168.
- Mulaw T. B., Kubicek C. P. and Druzhinina I. S. 2013. Novel endophytic Trichoderma spp. Isolated from Coffea arabica root are capable to controlling coffee tracheomycosis. Diversity 5: 750-766.
- Mysore, V. T.; Basavanna, M.; Monnanda, S. N.; Harishchandra, S. P.; Kukkundoor, R. K.; Ven, S.; Hunthrike, S. S. 2005. Endophytic fungal assemblages from inner and twig of Terminalia arjuna W. and A. (Combretaceae). World J. Microbiol. Biotechnol, 21, 1535-1540.
- Ohr H D; Munnecke D E; Bricker J L, 1973. The interaction of Armillaria mellea and Trichoderma spp. as modified by methyl bromide. Phytopathology, 63(8):965-973
- Paulitz T C. 2000 Population dynamics of biocontrol agents and pathogens in soil and rhizospheres. Eur J Plant Pathol; 106: 401-413.
- Ruppel, E. G., R. Baker, G. E. Harman, J. P. Hubbard, R. J. Hecker and I. Chet. 1983. Field tests of Trichoderma harzianum as a biocontrol agent of seedling disease in several crops and Rhizoctonia root rot of sugar beet. Crop Protection 2:399-408.
- Sivan, A., Y. Elad and I. Chet. 1984. Biological control of Pythium aphanidermatum by a new isolate of Trichoderma harzianum. Phytopathology 74:498-501.
Claims (13)
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Non-Patent Citations (16)
| Title |
|---|
| Abd-El-Moity TH, et al, 1981. Biological control of white rot disease of onion (Sclerotium cepivorum) by Trichoderma harzianum. Phytopathologische Zeitshrift 100, 29-35. |
| Abd-El-Moity TH, et al, 1983. Induction of new isolates of Trichoderma harzianum tolerant to © 2002 BSPP Plant Pathology (2002) 51, 735-745 744 J. P. Clarkson et al. fungicides and their experimental use for control of white rot of onion. Phytopathology 72, 396-400. |
| Benitez, T., et al. 2004. Biocontrol mechanism of Trichoderma strains. International Microbiol., 7: 249-260. |
| Chet. I. et al., 1981. Isolation and biocontrol potential of Trichoderma hamatum from soil naturally suppressive of Rhizoctonia solani. Phytopathology 71, 286-290. |
| Elad Y. 2000.Trichoderma harzianum T39 preparation for biocontrol of plant disease-control of Botrytis cinerea, Sclerotinia sclerotiorum and Cladosporium flavum. Biocontrol Sci Techn; 10: 499-507. |
| Elad, Y., et al. 1982.Control of Rhizoctonia solani in cotton by seed-coating with Trichoderma spp. spores. Plant Soil 66:279-281. |
| Hadar Y, et al. 1984. Evaluation of Trichoderma Koningii and T. hamatum from New York for biological control of seed rot caused by Pythium spp. Phytopathology, 74: 106-110. |
| Harman GE. 2000. Myths and dogmas of biocontrol: changes in perceptions derived from research on Trichoderma harzianum T-22. Plant Dis; 84(4): 377-393. |
| Harman, G.E. et al. 1983. Biological control of Pythium species. Seed Sci. Technol. 11:893-906. |
| Marois J.J., et al. 1982. Biological control of Verticillium wilt of eggplant in the field. Plant Disease 66, 1166-1168. |
| Mulaw T.B., et al. 2013. Novel endophytic Trichoderma spp. Isolated from Coffea arabica root are capable to controlling coffee tracheomycosis. Diversity 5: 750-766. |
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